Wang Na, Cheng Meimei, Zhang Xueyan, Wu Hongfei, Wu Huan, Cao Shijian, Wu Peng, Zhou An
The Experimental Research Center, Anhui University of Chinese Medicine, Hefei, China.
Anhui Province Key Laboratory of Chinese Medicinal Formula, Hefei, China.
Front Pharmacol. 2020 Oct 28;11:582390. doi: 10.3389/fphar.2020.582390. eCollection 2020.
This study was designed to investigate the therapeutic efficacy and underlying mechanisms of Gandou Decoction (GDD) in copper-laden hepatolenticular degeneration (HLD) model rats. In this study, high-performance liquid chromatography (HPLC) fingerprint analysis and eight representative active components were simultaneously measured for quality control of GDD. The therapeutic effect of GDD in HLD was studied by constructing a rat model of copper-laden HLD. The copper levels in the liver, serum, urine, and feces were quantified by atomic absorption spectrophotometry (AAS). Subsequently, UV-Vis spectrophotometry was used to study the coordination ability of copper ion (Cu) with six representative active components in GDD to explore its potential copper expulsion mechanism. Serological indexes including alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (AKP) were evaluated. Hepatic indicators including superoxide dismutase (SOD), glutathione (GSH), and the total antioxidant capacity (T-AOC) were determined. Moreover, the liver tissue was stained with hematoxylin-eosin to observe the histological changes. Thirty characteristic fingerprint peaks were used to assess the similarities among 10 samples and showed the similarity was >0.98, indicating a good correlation among the common peaks. Simultaneous quantification of eight markers in GDD was then performed to determine the consistency of quality. GDD could decrease the serum and hepatic copper levels by increasing the urinary and fecal copper content in copper-laden rats. Meanwhile, the results of UV-Vis absorption studies show that six representative active ingredients in GDD can coordinate with Cu, indicating that complexing copper removal may be a potential mechanism for GDD to play a role in copper removal. Serum hepatic enzyme markers AST, ALT, and AKP activities and antioxidant enzyme SOD, T-AOC activities, and GSH level in hepatic tissue showed the protection of GDD against liver injury induced by excessive copper. Additionally, the hepatoprotective effect of GDD was also evidenced by the results of the liver histological evaluation. This study suggested that GDD could reduce the serum and hepatic copper levels through promoting urinary and fecal copper excretion in copper-laden rats. At the same time, GDD could alleviate hepatic injury by inhibition of oxidative stress.
本研究旨在探讨肝豆汤（GDD）对肝豆状核变性（HLD）模型大鼠的治疗效果及潜在机制。本研究采用高效液相色谱（HPLC）指纹图谱分析并同时测定8种代表性活性成分，以对GDD进行质量控制。通过构建铜负荷HLD大鼠模型，研究GDD对HLD的治疗作用。采用原子吸收分光光度法（AAS）对肝脏、血清、尿液和粪便中的铜含量进行定量。随后，利用紫外可见分光光度法研究铜离子（Cu）与GDD中6种代表性活性成分的配位能力，以探索其潜在的排铜机制。评估血清学指标，包括丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）和碱性磷酸酶（AKP）。测定肝脏指标，包括超氧化物歧化酶（SOD）、谷胱甘肽（GSH）和总抗氧化能力（T-AOC）。此外，用苏木精-伊红对肝组织进行染色，观察组织学变化。用30个特征指纹峰评估10个样品之间的相似度，结果显示相似度>0.98，表明共有峰之间具有良好的相关性。随后对GDD中的8种标志物进行同时定量，以确定质量的一致性。GDD可通过增加铜负荷大鼠的尿铜和粪铜含量来降低血清和肝脏铜水平。同时，紫外可见吸收研究结果表明，GDD中的6种代表性活性成分可与Cu配位，表明络合排铜可能是GDD发挥排铜作用的潜在机制。血清肝酶标志物AST、ALT和AKP活性以及肝组织中抗氧化酶SOD、T-AOC活性和GSH水平表明，GDD对过量铜诱导的肝损伤具有保护作用。此外，肝脏组织学评估结果也证明了GDD的肝保护作用。本研究表明，GDD可通过促进铜负荷大鼠的尿铜和粪铜排泄来降低血清和肝脏铜水平。同时，GDD可通过抑制氧化应激减轻肝损伤。
